Accelerated solvent extraction of ochratoxin a from rice samples

In this paper, accelerated solvent extraction (ASE) for the analysis of ochratoxin A (OTA) is applied for the first time. Optimization of the method is given for the extraction of OTA from rice samples. Several parameters such as type of solvent, temperature, pressure, static time, and cell size were investigated thoroughly to find the optimal extraction conditions. The optimum ASE operating conditions were methanol as extraction solvent, 1500 psi, 40 degrees C, 5 min of static time, 50% flush volume, 60 s of purge, 1 cycle, and 11 mL cell size. The total extraction time was approximately 15 min. OTA was determined by liquid chromatography coupled with a fluorescence detector and confirmed by methyl ester derivatization. The analytical performance of the method was monitored with quality control parameters. Finally, the optimized method was used to evaluate 12 rice samples, 1 of which was positive with an OTA content of 4.17 ng/g. The proposed method offers the possibility of a fast and simple process to obtain a quantitative extraction of OTA.     

Assessment of insecticide resistance in eggs and neonate larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Spanish Cydia pomonella (L.) field populations have developed resistance to several insecticide groups. Diagnostic concentrations were established as the LC₉₀ calculated on a susceptible strain (S_Spain) for five and seven insecticides and tested on eggs and neonate larvae field populations, respectively. The three most relevant enzymatic detoxification systems (mixed-function oxidases (MFO), glutathione S-tranferases (GST) and esterases (EST)) were studied for neonate larvae.In eggs, 96% of the field populations showed a significantly lower efficacy when compared with the susceptible strain (S_Spain) and the most effective insecticides were fenoxycarb and thiacloprid. In neonate larvae, a significant loss of susceptibility to the insecticides was detected. Flufenoxuron, azinphos-methyl and phosmet showed the lowest efficacy, while lambda-cyhalothrin, alpha-cypermethrin and chlorpyrifos-ethyl showed the highest. Biochemical assays showed that the most important enzymatic system involved in insecticide detoxification was MFO, with highest enzymatic activity ratios (5.1-16.6 for neonates from nine field populations). An enhanced GST and EST activities was detected in one field population, with enzymatic activity ratios of threefold and fivefold for GST and EST, respectively, when compared with the susceptible strain. The insecticide bioassays showed that the LC₉₀ used were effective as diagnostic concentrations. Measures of MFO activity alongside bioassays with insecticide diagnostic concentrations could be used as tools for monitoring insecticide resistance in neonate larvae of C. pomonella.     

A stochastic optimization method to estimate the spatial distribution of a pathogen from a sample

Information on the spatial distribution of plant disease can be utilized to implement efficient and spatially targeted disease management interventions. We present a pathogen-generic method to estimate the spatial distribution of a plant pathogen using a stochastic optimization process which is epidemiologically motivated. Based on an initial sample, the method simulates the individual spread processes of a pathogen between patches of host to generate optimized spatial distribution maps. The method was tested on data sets of Huanglongbing of citrus and was compared with a kriging method from the field of geostatistics using the well-established kappa statistic to quantify map accuracy. Our method produced accurate maps of disease distribution with kappa values as high as 0.46 and was able to outperform the kriging method across a range of sample sizes based on the kappa statistic. As expected, map accuracy improved with sample size but there was a high amount of variation between different random sample placements (i.e., the spatial distribution of samples). This highlights the importance of sample placement on the ability to estimate the spatial distribution of a plant pathogen and we thus conclude that further research into sampling design and its effect on the ability to estimate disease distribution is necessary.     

Genetic variation and genotype-environment interactions in short rotation <Emphasis Type="Italic">Populus</Emphasis> plantations in southern Europe

Growth and production in the first year, as additional selection criteria, were assessed for nine poplar clones to be used as short rotation woody crops (SRWC) in the production of biomass for energy purposes. In order to identify the most promising clones in terms of growth and yield and also to assess their stability, trials were established at different locations in Spain. The majority of these clones, which form part of the European list of base materials, have frequently been used in plantations aimed at timber production but not for biomass in Mediterranean conditions. Others, such as those selected in Italy specifically for biomass production (currently provisionally admitted), are being tested for the first time under different soil and climatic conditions in Southwest Europe. The early selection of clones for rapid juvenile growth provides a valuable additional input to the clonal selection process, especially where very short rotations are desired (no more than 3 years). In any case, determining clonal stability in terms of growth is of great use not only when deciding on the clones to be used in plantations but also when developing breeding programs. ANOVA and Genotype plus Genotype × Environment (GGE) biplot analyses were used to analyse the growth and stability of the clones, which were then ranked according to mean performance and stability. Differences were detected between clones as well as between the different environments tested. The biplot analysis allowed different groups of clones to be identified according to their performance and degree of interaction displayed, thus providing useful information for the selection process. The production of aboveground biomass in the first vegetative period ranged from 1.7 to 8.0 Mg DM ha⁻¹ at the different sites. ‘Monviso’, ‘Guardi’, ‘AF2’ and ‘2000 verde’ were the most productive clones whereas ‘Unal’, ‘Pegaso’ and ‘USA 49-177’ were the least productive. The stability analysis identified ‘AF2’, ‘Guardi’, ‘I-214’ and ‘MC’ as more stable clones while ‘Monviso’, ‘2000 verde’, ‘Unal’, ‘Pegaso’ and ‘USA 49-177’ were found to be specifically adapted to certain environments. This implies that where information on site conditions is not available, the ‘AF2’ and ‘Guardi’ clones offer greater assurance of successful establishment and higher initial growth. The growth of ‘Monviso’ ‘2000 verde’ ‘Unal’ ‘Pegaso’ and ‘USA 49-177’ clones is highly dependent on site conditions during the establishment phase. Similarly, the SH (Shore Henares river) and LT (La Tallada) sites were identified as the most highly discriminative environments for the set of clones while CS (Cubo de la Solana) and AR (Atarfe) were identified as those where performance levels were average.     

Simultaneous exposure of forest trees to pollutants and climatic stress

This paper describes a facility designed to simultaneously generate complex climatic and pollutant conditions for research into the effects of climate and air pollution on plants. The system, commissioned in 1985, consists of a set of 4 environmental chambers which are each equipped with four identical subchambers. These 4 subchambers are subjected to identical climatic conditions, but are equipped with independent gas supplies. The chambers are designed to operate within a temperature range of -20 to +40° C at a relative humidity of 20 to 95 % with an illumination system providing high light intensities at a spectrum very close to sunlight. A computer controlled fumigation system allows the injection of various gases singly and in combination at concentrations between 0.005 to l,000 ppm (SO₂, NO_x, O₃, CO₂, PAN).     

Einfluß langjähriger Düngung mit verschiedenen N-Formen auf die Phosphat-Dynamik einer Ackerbraunerde

Influence of long-term application of different nitrogen sources on the dynamics of phosphate in an arable brown earth In a long-term field experiment parameters of the soil phosphate status were determined after 53 years. Soil characteristics of the arable brown earth are: sandy silt loam; pH 5,9–6,6; total carbon 0,9%; CEC 13–15 meq/100 g soil; effective field capacity 160 mm unto 150 cm. The following results were obtained: Total P content was slightly higher in the plots without mineral N and with farm manure (90 and 93 mg P/100 g soil resp.) than in the plots with mineral N supply (Ø 85 mg P/100 g soil). Application of farm yard manure and ammonium sulfate caused higher proportions of ?Fe and Al phosphates”? (56–57% of inorganic phosphate) whereas calcium cyanamide, calcium nitrate and ammonium sulfate with annual liming led to higher a content of Ca-phosphate (52–56% of inorganic phosphate). When pH was brought to 6,5 by liming, P uptake of rye grass and green oats/green rape-seed in a pot experiment depended on the soil content of ?Fe- and Al-phosphates”?. Without liming, P uptake was lower in the plot with ammonium sulfate, however, in the calcium cyanamide plot P uptake was higher than expected from the content of ?Fe- and Al-phosphate”?.     

Ionic interactions and salinity affect monoterpene and phenolic diterpene composition in rosemary (Rosmarinus officinalis)

In this study, we evaluated how increased cation supply can alleviate the toxic effects of NaCl on plants and how it affects essential oils (EOs) and phenolic diterpene composition in leaves of rosemary (Rosmarinus officinalis L.) plants grown in pots. Two concentrations of the chloride salts KCl, CaCl₂, MgCl₂, and FeCl₃ were used together with 100 mM NaCl to study the effects of these nutrients on plant mineral nutrition and leaf monoterpene, phenolic diterpene, and EO composition. The addition of 100 mM NaCl, which decreased K⁺, Ca²⁺, and Mg²⁺ concentrations with increasing Na⁺ in leaves, significantly altered secondary metabolite accumulation. Addition of MgCl₂ and FeCl₃ altered leaf EO composition in 100 mM NaCl–treated rosemary plants while KCl and CaCl₂ did not. Furthermore, addition of CaCl₂ promoted the accumulation of the major phenolic diterpene, carnosic acid, in the leaves. The carnosol concentration was reduced by the addition of KCl to salt‐stressed plants. It is concluded that different salt applications in combination with NaCl treatment may have a pronounced effect on phenolic diterpene and EO composition in rosemary leaves thus indicating that ionic interactions may be carefully considered in the cultivation of these species to achieve the desired concentrations of these secondary metabolites.     

A comparative study of apple and pear tree water consumption measured with two weighing lysimeters

A five-year experiment (2002–2006) was conducted to determine apple (cv ‘Golden Smoothee’) and pear (cv ‘Conference’) crop coefficients (Kc) using two large weighing lysimeters. Daily reference evapotranspiration (ETo) and crop evapotranspiration (ETc) were evaluated. Midday canopy light interception of both crops planted in hedgerows, 4 × 1.6 m, was determined on a weekly basis from bud-break until leaf fall from year 2002 (fourth after planting) to year 2006 of both plantations. Relationships between canopy light interception and calculated Kc (ETc/ETo) were evaluated from bud-break until harvest. There were differences in Kc values between apple and pear trees. When daily Kc values from bud-break until harvest were adjusted to hyperbolic functions each year, adjusted curves for pear trees were very similar regardless of year (maximum Kc around 1.0). In apple trees, the maximum values of Kc increased over time, from 0.49 in 2002 to 1.04 in 2006. Midday light interception in both apple and pear trees increased during the 5 years of experiment from 29.0 to 45.6% in apples and from 27.5 to 41.6% in pears in midsummer. Although there was a significant positive correlation between midday canopy light interception and Kc in apple and pear trees, in different times within a specific year, these relationships were different between crops. While the apple data fitted into the same equation regardless of the year, different equations were needed to fit the pear data in different years. This discrepancy may have been related to differences in the canopy properties between apple and pear trees. Pear canopies had higher porosity than apple canopies and thus improved light penetration. Apple trees were more vigorous and produced taller and denser canopies. Pear Kc values were greatly influenced by the evaporative demands of different years and consequently differences in midday canopy light interception did not adequately reflect the differences in Kc across the two species.     

Fruit thinning in ‘Conference’ pear grown under deficit irrigation: Implications for fruit quality at harvest and after cold storage

Fruit thinning in pear is feasible for mitigation of water stress effects. However, it is not well known how fruit quality at harvest and after cold storage is affected by pre-harvest water stress. Even less is known about the effects of fruit thinning on quality under these circumstances. To elucidate these, we applied deficit irrigation (DI) and fruit thinning treatments to ‘Conference’ pear over the growing seasons of 2008 and 2009. At the onset of Stage II (80 and 67 days before harvest in 2008 and 2009, respectively), two irrigation treatments were applied: full irrigation (FI) and DI. FI trees received 100% of crop evapotranspiration (ETc). DI trees received no irrigation during the first three weeks of Stage II to induce water stress, but then received 20% of ETc to ensure tree survival. From bud-break until the onset of Stage II and during post-harvest, FI and DI trees received 100% of ETc. Each irrigation treatment received two thinning levels: no thinning leaving commercial crop load (∼180 fruits tree⁻¹), and hand-thinning at the onset of Stage II leaving a light crop load (∼85 fruits tree⁻¹). Under commercial crop loads, DI trees were moderately water-stressed and this had some positive effects on fruit quality. DI increased fruit firmness (FF), soluble solids concentrations (SSC) and acidity at harvest while no changes were observed in fruit maturity (based on ethylene production). Differences in FF and acidity at harvest between FI and DI fruit were maintained during cold storage. DI also reduced fruit weight loss during storage. But fruit size was reduced under DI. Fruit thinning under DI resulted in better fruit composition with no detrimental effect on fresh-market yield compared to un-thinned fruit. Fruit size at harvest and SSC values after five months of cold storage were higher in fruit from thinned trees than fruit from un-thinned trees. Fruit thinning increased fruit ethylene production, indicating advanced maturity. This may lead to earlier harvest which is desirable in years with impending drought. Fruit thinning is therefore a useful technique to enhance pear marketability under water shortage.     

Factors associated with routine mass antimicrobial usage in fattening pig units in a high pig-density area

One hundred and seven fattening pig units randomly selected in Catalonia (Spain) were surveyed during 2001-2003, in order to determine the frequency of the prophylactic use of antimicrobials and mass treatments against respiratory and enteric disease outbreaks. Logistic regression and regression tree analysis were done to determine the factors associated with the on-farm mass antimicrobial usage. Fifty-eight per cent of the surveyed farms used mass antimicrobial prophylaxis; among them, 39% used two or more antimicrobials. The more frequently administered drugs were colistin, beta-lactams and tetracyclines. Fattening units from strict finishing farms were more likely to use antimicrobials than farrow-to-finish farms (Odds Ratio = 11.7) and farms with changing facilities were less likely to use these compounds (OR = 0.17). In the event of a respiratory disease outbreak, most farms applied mass treatment and 46% used two or more compounds, with tetracyclines, beta-lactams and sulphonamides being the most popular ones. Mass treatment with several compounds in case of respiratory problems was significantly related to the use of growth promoters on the farm (OR = 4.3). If an outbreak of an enteric disease occurred, most farms resorted also to a mass treatment and 48% used two or more drugs. The most frequent antimicrobial agent for the treatment of enteric disease was colistin. Fattening farms (OR = 6.9) and operations that perform routine antimicrobial prophylaxis (OR = 2.0) were more likely to use more than one antimicrobial agent for enteric diseases.